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Research article

High doses of siRNAs induce eri-1 and adar-1 gene expression and reduce the efficiency of RNA interference in the mouse

Jie Hong, Zhikang Qian, Shuiyuan Shen, Taishan Min, Chang Tan, JianFeng Xu, Yingchun Zhao, Weida Huang
Biochemical Journal Sep 15, 2005, 390 (3) 675-679; DOI: 10.1042/BJ20050647
Jie Hong
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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Zhikang Qian
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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Shuiyuan Shen
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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Taishan Min
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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Chang Tan
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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JianFeng Xu
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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Yingchun Zhao
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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Weida Huang
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
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  • For correspondence: whuang@fudan.edu.cn
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This article has a commentary. Please see:

  • Dampening the silencing effect of RNA interference in mammals

Abstract

RNAi (RNA interference) is a gene-silencing mechanism that is conserved in evolution from worm to human and has been a powerful tool for gene functional research. It has been clear that the RNAi effect triggered by endogenous or exogenous siRNAs (small interfering RNAs) is transient and dose-dependent. However, there is little information on the regulation of RNAi. Recently, some proteins that regulate the RNA-silencing machinery have been identified. We have observed in previous work that the expression of target genes rebounds after being suppressed for a period of time by siRNAs. In the present study, we used secretory hepatitis B virus surface antigen gene as a reporter and compared its expression level in cell culture and mice challenged by different doses of siRNAs. A quicker and higher rebound of gene expression was observed in mice tail-vein-injected with higher doses of siRNA, and the rebound was associated with an increase in the mRNA level of meri-1 (mouse enhanced RNAi) and adar-1 (adenosine deaminase acting on RNA) genes encoding an exonuclease and RNA-specific adenosine deaminase respectively. Down-regulation of meri-1 by RNAi enhanced the sensitivity and efficiency of siRNA in inhibiting the expression of hepatitis B virus surface antigen. These results indicate that RNAi machinery may be under negative regulation, through the induction of a series of genes coding for destabilizing enzymes, by siRNAs introduced into the cell, and also suggest that a suitable amount of siRNA should be used for research or therapeutic applications.

  • Escherichia-coli-expressed and enzyme-digested RNA (esiRNA)
  • hydrodynamic injection
  • mouse enhanced RNA interference gene (meri-1)
  • RNA interference (RNAi)
  • RNA interference regulation
  • small interfering RNA (siRNA)

Abbreviations: Adar, adenosine deaminase acting on RNA; CHO, Chinese-hamster ovary; dsRNA, double-stranded RNA; eri, enhanced RNA interference; esiRNA, Escherichia-coli-expressed and enzyme-digested siRNA; HBsAg, type B hepatitis virus surface antigen; HBVP, type B hepatitis virus polymerase; meri-1, mouse eri-1; NP, nucleoprotein; RDRP, RNA-dependent RNA polymerase; RISC, RNA-induced silencing complex; RNAi, RNA interference; RT, reverse transcription; siRNA, small interfering RNA

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September 2005

Volume: 390 Issue: 3

Biochemical Journal: 390 (3)
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High doses of siRNAs induce eri-1 and adar-1 gene expression and reduce the efficiency of RNA interference in the mouse
Jie Hong, Zhikang Qian, Shuiyuan Shen, Taishan Min, Chang Tan, JianFeng Xu, Yingchun Zhao, Weida Huang
Biochemical Journal Sep 2005, 390 (3) 675-679; DOI: 10.1042/BJ20050647
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High doses of siRNAs induce eri-1 and adar-1 gene expression and reduce the efficiency of RNA interference in the mouse
Jie Hong, Zhikang Qian, Shuiyuan Shen, Taishan Min, Chang Tan, JianFeng Xu, Yingchun Zhao, Weida Huang
Biochemical Journal Sep 2005, 390 (3) 675-679; DOI: 10.1042/BJ20050647

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Keywords

Escherichia-coli-expressed and enzyme-digested RNA (esiRNA)
hydrodynamic injection
mouse enhanced RNA interference gene (meri-1)
RNA interference (RNAi)
RNA interference regulation
small interfering RNA (siRNA)

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